Method for constructing eukaryotic expression vector by virtue of designing multiple cloning sites (MCS)

Abstract

The invention belongs to the technical field of biology, and particularly relates to a method for constructing a eukaryotic expression vector by virtue of designing multiple cloning sites (MCS). The eukaryotic expression vector is obtained by connecting restriction enzyme cutting sites without eukaryotic gene promoters, tanscription termination regulatory sequences, selection marker genes, exogenous target genes and primary skeleton vector sequences together, and then cloning the restriction enzyme cutting sites to an MSC-free primary skeleton vector, and finally using a DNA (Deoxyribose Nucleic Acid) recombinant technology to insert the eukaryotic gene promoters, the tanscription termination regulatory sequences, the selection marker genes and the exogenous target genes into the skeleton vector containing a target restriction enzyme cutting site sequence. The method is flexible and practical, is suitable for the construction of all simple and complete eukaryotic expression vectors, and can be used for overcoming the defect that the inherent restriction enzyme cutting sites on the primary skeleton vector cannot meet requirements; and after the proper restriction enzyme cutting sites are determined, and the subsequent vector construction work is fast and efficient, so that the process of the research on a laboratorial eukaryotic gene function is accelerated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing eukaryotic expression vectors by designing multiple cloning sites. Background technique [0002] With the completion of the genome project of human beings and different species, the role of gene manipulation technology in life science research is becoming more and more important. The construction of eukaryotic expression vectors is the core of gene manipulation technology. At present, the construction technology of eukaryotic expression vectors has been developed rapidly and is widely used in laboratories, biopharmaceuticals, transgenic animals and plants and other fields. [0003] The key elements for constructing eukaryotic expression vectors are gene promoters, transcription termination regulatory sequences, screening marker genes and exogenous target genes from the same or similar eukaryotic cells. The current common practice is to use the p...

AI Technical Summary

Problems solved by technology

Due to the limitation of the limited multiple cloning sites on the backbone vector, suitable restriction sites are often not found, which seriously affects the efficiency of constructing eukaryotic expression vectors

When constructing a double expression cassette vector, more suitable restriction sites for sorting are needed. Using existing methods, the problem of restricting construction efficiency will become more prominent

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